Comparison of use of cerebrospinal fluid, serum, and throat swab specimens in diagnosis of enteroviral acute neurological infection by a rapid RNA detection PCR assay.

A PCR assay for detection of enterovirus RNA in multiple specimen types from patients with neurological infections was evaluated. Combined PCR assay of cerebrospinal fluid and serum (systemic specimens) was more sensitive than assaying either specimen alone in children but not in adults. Compared with PCR in systemic specimens, detection of enterovirus RNA in throat swabs showed a sensitivity of 62.

[PCR value in the diagnosis of feto-placental human parvovirus B19 hydrops fetalis: apropos of 10 cases].

Human parvovirus B19 primary infection during pregnancy is responsible for 27% of non autoimmune hydrops fetalis. Parvovirus B19 antigen detection and parvovirus B19 IgM and IgG antibody determination using enzyme immunoassays are not reliable for diagnostic purposes and lack of specificity. Parvovirus B19 DNA detection in amniotic fluid, fetal blood, ascitic fluid, and fetal biopsies or placenta specimens seems to be the best method for the diagnosis.

Procalcitonin as a marker of bacterial sepsis in patients infected with HIV-1.

Procalcitonin (ProCT) is a recently described marker of severe sepsis. It was decided to assess the value of proCT as a marker of secondary infection in patients infected with HIV-1. ProCT plasma levels were measured by immunoluminometric assay in a prospective study in 155 HIV-infected individuals: 102 asymptomatic and 53 with lever or suspected secondary infections.

Experimental CVB3-induced chronic myocarditis in two murine strains: evidence of interrelationships between virus replication and myocardial damage in persistent cardiac infection.

In order to analyse the relationships between enteroviral replication and the myocardial damage at the onset of chronic cardiac infection, 2 mouse strains with different degrees of immunological competence (NMRI nu/nu, DBA/2) were infected by a myocarditic Coxsackie virus B3 (CVB3-M1) variant. At 31 days post-inoculation, plaque-forming assay, polymerase chain reaction (RT-PCR), and immunohistochemistry were carried out for detecting viruses and viral components in the myocardium. The virological findings were related to histopathological changes in the myocardium as well to the dilatation of both cardiac ventricles.

Detection of coxsackie B virus RNA sequences in whole blood samples from adult patients at the onset of type I diabetes mellitus.

Enteroviruses may be linked to insulin-dependent diabetes mellitus (IDDM). The prevalence of enteroviral (EV) infection at onset of adult IDDM was investigated by detection of specific EV sequences in peripheral blood using a reverse transcription and a seminested polymerase chain reaction (seminested RT-PCR). EDTA-treated whole blood samples taken from 12 newly diagnosed IDDM patients with ketosis or ketoacidosis were examined.

An interferon-gamma (IFN-gamma) based whole blood assay to detect T cell response to antigens in HIV-1 infected patients.

Recently it has been reported that cytokine production by T cells in response to antigens may be more sensitive test than lymphoproliferation. T cell reactivities to antigens is usually performed on isolated PBMCs, however whole blood is being used frequently for cytokine production studies. A whole blood assay is described to measure T cell mediated immune responses to HIV-1 and recall antigens.

A macrophage-like cell model for testing anti-CMV drugs.

Monocyte/macrophage cells can be infected with human cytomegalovirus (HCMV) and could be a reservoir and a vehicle for virus spread in infected patients. The monocytic-like cell line THP-1 differentiated along the monocyte pathway with a phorbol ester (phorbol 12-myristate 13-acetate, PMA) for 24h was infected with cell free-supernatant of Towne CMV-strain infected fibroblasts MRC-5. We designed experiments to study the replication of HCMV in PMA treated THP-1 cells, exposed to DHPG (Cymevan, ganciclovir, Syntex).

Production of TNFalpha and IL-6 by activated whole blood from HIV-1 infected patients detected by a one-stage procedure: relationship with the phenotype of HIV-1 isolates.

Diluted whole blood (WB) culturing may be the most appropriate milieu in which to study cytokine production in vitro. We tested TNFalpha and IL-6 production using small volumes of WB (25 microl) from HIV-1 positive patients with a one-step procedure that combines WB stimulation with LPS, PHA and cytokine measurement. We studied 49 patients without secondary infection or at distance of secondary infection staged according to the 1993 classification of the CDC and 12 healthy seronegative subjects.

Multicenter evaluating of a commercially available PCR assay for diagnosing enterovirus infection in a panel of cerebrospinal fluid specimens.

Thirteen laboratories participated in blind tests of a panel of 20 coded cerebrospinal fluid specimens (7 uninfected samples, 3 samples infected with 1 50% tissue culture infective dose [TCID50]/0.1 ml [nonenterovirus strains], and 10 samples infected with 10, 1, or 0.1 TCID50/0.

Enhanced TNF alpha production by monocytic-like cells exposed to dengue virus antigens.

The studies indicating the importance of TNF alpha in dengue virus infection have led us to determine whether monocyte-like cells produce TNF alpha exposure after dengue virus. The supernatant fluids of mosquito cells (AP61) infected with dengue virus (DV) type 1 and DV type 3 were harvested 7 days post-infection and clarified. DV inactivation was performed in the presence of betapropiolactone that preserves antigenicity of viruses.

Virucidal activity of pure singlet oxygen generated by thermolysis of a water-soluble naphthalene endoperoxide.

Using the water-soluble naphthalene carrier of singlet oxygen NDPO2, we have shown that pure singlet oxygen is able to inactivate enveloped viruses (human immunodeficiency virus type 1, herpes simplex virus type 1, cytomegalovirus, vesicular stomatitis virus), but has no effect on non-enveloped viruses (adenovirus and poliovirus 1). These results are related to the experiments on photoinactivation of viruses by hydrophobic photosensitizers (merocyanine 540, hypericin, phthalocyanines, hematoporphyrin and benzoporphyrin derivatives) and they strengthen the hypothesis that singlet oxygen plays a predominant role in this process.

Rapid detection of enterovirus in clinical specimens using PCR and microwell capture hybridization assay.

A rapid detection method of enteroviral RNA in clinical samples using PCR and a microwell capture hybridization assay is described. PCR products were labelled directly by digoxigenin-dUTP during the amplification step. The labelled amplicons were hybridized with a biotinylated oligo-probe and captured on commercially available test microwells coated with streptavidin.

Plasma levels of sTNFR p75 and IL-8 in patients with HIV-1 infection.

We determined the degree of activation of the TNF alpha system and the levels of IL-8 in HIV-1 infection. TNF alpha is one of the most potent agents for the induction of IL-8. TNF alpha may be cleared rapidly from the circulation, however soluble tumor necrosis factor receptor (sTNFR) p75 which is more stable may reflect the degree of activation of the TNF alpha system.

High plasma level of soluble tumor necrosis factor receptor type II (sTNFRII) in asymptomatic HIV-1-infected patients.

Plasma concentrations of soluble receptors for tumor necrosis factor type II (sTNFRII) and CD4+ lymphocyte counts were determined in patients with HIV-1 infection grouped according to the 1993 classification of the CDC. Compared with healthy controls (mean +/- SD = 2.83 +/- 0.

[Polyvalent hyperimmune immunoglobulins prevent the infection of monocytic type cells by human cytomegalovirus].

Monocyte/macrophage cell types can be infected with Human Cytomegalovirus (HCMV) could be a reservoir and a vehicle for virus spread in infected patients. We developed a model to study the effects of antiviral molecules on these cells. The monocytic-like cell line THP-1 and the human diploïd cells MRC-5 were used.

TNF alpha production by whole blood from HIV-1 infected patients.

Production of cytokines by immunocompetent cells in vitro may be assessed after stimulation with polyclonal activators. Because it mimics the natural environment, diluted whole blood (WB) culture may be the most appropriate milieu in which to study cytokine production in vitro. We tested TNF alpha production by small volume of whole blood (25 microliters) from HIV-1 positive patients by using a one-step procedure that combines WB stimulation with LPS and PHA and cytokine measurement.

Sensitization of AIDS related non-Hodgkin's B lymphoma cell lines to cytotoxic drugs toxins by interferon-gamma.

AIDS-related lymphomas (ARL) progressively become resistant to conventional chemotherapy. We have developed three B cell lines from tumor biopsies of AIDS patients with non-Hodgkin's lymphoma (NHL). The ARL cell lines were shown to be resistant to a panel of cytotoxic cytokines, toxins and drugs such as tumor necrosis factor, diphteria toxin, ricin, adriamycin, cis-platinum and anti-Fas antibody.

Coxsackievirus B3-induced chronic myocarditis in mouse: use of whole blood culture to study the activation of TNF alpha-producing cells.

The pathogenesis of CVB3-induced chronic myocarditis remains unknown. Activated monocytes and macrophages may maintain ongoing inflammation during a persistent CVB3 infection and possibly represent the major mechanism leading to chronic myocarditis. We decided to study the activation status of cells by studying TNF alpha secretion in vitro using whole blood culture in CVB3-induced murine chronic myocarditis.